OBJECTIVE: To elongate the proliferation life-span of human umbilicus vein endothelial cell (HUVEC). METHODS: We synthesized the human telomerase reverse transcriptase mRNA (hTERT mRNA) by in vitro transcription, then transferred the hTERT mRNA into HUVEC in quicent stage by lipofect introduction. RESULTS: Telomerase expressed transiently in HUVEC, and the cell life-span was elongated for 7 population doublings. CONCLUSION: Telomerase can be reconstructed controllably and transiently in HUVEC by hTERT mRNA introduction, this method has the potential to be used to elongate the lifespan of cells cultured in vitro.
Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes， the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P＜0.05). ②The expression of c-met mRNA was correlated with both the metastatic lymph nodes and tumor size. ③There was no significant relationship between the expression of c-met mRNA and the expressions of ER, PR and c-erbB-2. ④Dounching operative field with 5-FU and distilled water could decrease the expression of c-met mRNA.Conclusion The proto-oncogene c-met mRNA may be an ideal and specific marker for dectecting micrometastasis of breast cancer. In addition, it also suggests that the examination of c-met mRNA in the axillary drainage by RT-PCR assay could detect the micrometastasis in axillary lymph nodes much easier and more accurately than routine pathological method.
Objective To investigate the effects of expression of TNFα mRNA on glucose uptake in both the liver and skeletal muscle after endotoxemia. Methods In the mice with intraperitoneal injection of lipopolysaccharide (LPS), the changes of TNFα level of plasma and uptake of 2-deoxyglucose (2-DG) in the isolated soleus muscle and hepatic tissues were determined, then the reinstatement of glucose uptake by injecting TNF-McAb for 3 days was also observed. In addition, changes of TNFα mRNA expression of liver were evaluated. Results The expression of TNFα mRNA in the liver showed markedly increased in the first 3 hours post endotoxemia and remaind high for 3 days, and the plasma TNFα level paralleled with TNFα mRNA expression of liver also was elevated. The basal uptake of 2-DG both in muscle and liver were markedly increased, but the stimulated 2-DG uptake with insulin was greatly reduced as compared with the control. In addition, these abnormalities of 2-DG uptake can be partially corrected by neutralization of the circulatory TNFα by administration of TNF-McAb. Conclusion The disorders of glucose uptake of the liver and the muscle due to the overexpression of TNFα mRNA and elevated circulatory TNFα level may be the mechanism of insulin resistance after endotoxemia.
【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
Objective To investigate the percentage of CD4 + CD25 + Treg cells and expression of Foxp3 mRNA in asthmatic patients and the impacts of inhaled steroids.Methods The percentages of CD4 +CD25 + Treg cells was assayed by flow cytometry and the expression of Foxp3 mRNA was detected by RT-PCR in peripheral blood mononuclear cells from the patients with chronic persistent asthma before and after steroids inhalation in comparison with healthy control. The forced expired volumin one second/predicted value( FEV1% pred) and peak expired flow( PEF) were measured by spirometry. Results The level of CD4 + CD25 + Treg cells and the expression of Foxp3 mRNA were lower in asthmatics before steroids treatment than those in control ( P lt; 0. 05) which were increased significantly after steroids treatment ( P lt; 0. 05) .FEV1% pred and PEF were declined significantly than those in control but improved markedly after treatment ( P lt; 0. 05) . Conclusions The insufficiency of amount and function of immue-suppressive CD4 + CD25 +Treg cells may play a role in the pathogenesis of asthma. Inhaled steroids can improve the lung function of asthmatics by upregulating the level of CD4 + CD25 + Treg and Foxp3.
Objective To investigate the relationship between gene expression of endothelin-3 (ET-3） and inflammation of acute pancreatitis （AP） in rats. Methods Fifty-four rats were divided randomly into 4 groups: sham operation group, AP group, arterial injection group and vein injection group. AP was induced by reverse intra-bile duct infusion 4.5% sodium taurocholate, treated with low dose dopamine 〔5 μg/（kg·min）〕 by injecting arterial or tail vein. Rats were sacrificed at 1， 6 and 24 h after the induction of AP. The mRNA expression of ET-3 was evaluated by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and pathological changes was observed in rats. Results Expression of ET-3 mRNA could be detected from 1 up to 24 h after the induction of pancreatitis. Expression of ET-3 mRNA of sham operation group was decreased significantly compared with other three groups. Expression of ET-3 mRNA showed a significant decrease by arterial injection dopamine than that by tail vein (P＜0.05， P＜0.01). The pathologic score in AP group was the highest, vein injection group was the next one, and score in sham operation group was the lowest. Conclusion There are significant relationship between inflammation of AP and expression of ET-3 mRNA. Dopamine administration by arterial injection is more effective than that by tail vein injection.
【Abstract】ObjectiveTo investigate the expression of Tob mRNA in human colorectal cancer tissues, and their corresponding paracancerous normal tissues which was 10 cm above the tumor and pathologically proved and to explore the role of Tob mRNA in the pathogenesis of colorectal cancer. MethodsQuantitative real time RTPCR was used to detect the expression of Tob mRNA in 31 colorectal cancers. ResultsCompared with paracancerous tissue, the expression of Tob mRNA in colorectal cancer tissues was significantly increased. Moreover, the expression levels of Tob in Dukes A, B, C, D were 1.146±0.067, 1.120±0.073, 1.052±0.020 and 1.047±0.010 respectively. Analyzed by oneway ANOVA, there were significant differences in expression of Tob in different Dukes stage. ConclusionThe upregulation expression of Tob mRNA may be closely associated with tumorigenesis of colorectal carcinoma.
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.
Objective To detect expression of CK20 mRNA in peripheral blood of patients with colorectal carcinoma and its clinical significance. Methods Using the reverse transcriptase-polymerase chain reaction (RT-PCR),CK20 mRNA expression was examined in peripheral blood from 42 patients with colorectal carcinoma before and after operation, 20 healthy volunteers, 20 fresh colorectal carcinoma samples. Results The positive expression rates of CK20 mRNA were 45.24%(19/42) and 33.33%(14/42) before and after operation in 42 colorectal carcinoma patients respectively. All 20 fresh colorectal carcinoma samples revealed expression of CK20 mRNA, but the 20 normal blood samples did not. Conclusion The detection of CK20 mRNA in peripheral blood is helpful to early diagnose, assess the prognosis and make a correct treatment of colorectal carcinoma.
ObjectiveTo evaluate the effects of high intensity focused ultrasound (HIFU) on hepatic cancer cells spreading in blood.Methods AFP mRNA in peripheral blood of 19 patients with hepatocellular carcinoma was detected before and after HIFU therapy by RTPCR.Results①Before HIFU therapy, 11 of 19 cases were AFP mRNA positive (57.9%), while the control group were all negative. AFP mRNA was correlated with some clinical parameters such as serum AFP level, tumor size, portal vein embolism and extrahepatic metastasis (P＜0.05). ②In 8 cases with preoperative AFP mRNA negative, only 2 cases became AFP mRNA positive immediately after therapy, and one of the 2 cases became negative again after 72 hours. One week after HIFU therapy, the AFP mRNA positive rate (31.6%) was much lower than the preoperative positive rate (57.9%), but there was no statistical significance (Pgt;0.05). ③After one week of HIFU therapy, the AFP mRNA positive rate of the group with tumor size less than 8 cm was much lower than that of tumor size larger than 8 cm (P＜0.05). ConclusionHIFU may reduce the spreading of hepatocellular carcinoma cells in blood. It is effective for patients with tumor size less than 8 cm.